Figure 1.

SLC10A4 co-localises with mMCP-6 and lack of SLC10A4 does not interfere with the storage of mMCP-6 inside the mast cell granules. (A) Wild type and Slc10a4 ^−/− BMMCs were immunostained with anti-SLC10A4 antibody (green), counterstained with DAPI (blue) and analysed by immunofluorescence microscopy. (B) Wild type BMMCs were stained with anti-SLC10A4 (green) and anti-mMCP-6 (red) antibodies and counterstained with DAPI to visualize granular staining. (B,C) The majority of the mast cell granules demonstrated co-localization (yellow) of mMCP-6 and SLC10A4 as indicated in the Venn diagram (analysis of 112 wild type BMMCs). (D) Representative pictures of wild type (WT) and Slc10a4 ^−/− BMMCs immunostained for mMCP-6 (red) and counterstained with DAPI (blue). As a negative control, BMMCs were stained with the secondary anti-goat antibody alone (control). (E) Quantitative western blot analysis of mMCP-6 expression in BMMCs derived from WT and Slc10a4 ^−/− mice. The quantification analyses were performed by western blot on cell lysate and mMCP-6 expression levels were normalised to the β-actin amount in the same sample. The data shown are presented as mean ± SEM from six samples obtained from three independent western blots. No significant difference (p = 0.18) was found using a standard unpaired Student’s t-test. (F) WT and Slc10a4 ^−/− bone marrow cells were cultured with SCF and IL-3 to obtain BMMCs. Samples from the indicated days after the start of the cultures in vitro (days in vitro; DIV) were cytospun, stained with May-Grünwald/Giemsa and evaluated as positive or negative for cells with metachromatic staining. These data represent pooled data from two mice per genotype where the percentage of cells with metachromatic staining were calculated from the individual cell cultures based on three individual cytospins per data point. Scale bars, 10 µm.